@article{b679f05d42c2475da87a7c0fd8e30124,
title = "An isomorphous replacement method for efficient de novo phasing for serial femtosecond crystallography",
abstract = "Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) holds great potential for structure determination of challenging proteins that are not amenable to producing large well diffracting crystals. Efficient de novo phasing methods are highly demanding and as such most SFX structures have been determined by molecular replacement methods. Here we employed single isomorphous replacement with anomalous scattering (SIRAS) for phasing and demonstrate successful application to SFX de novo phasing. Only about 20,000 patterns in total were needed for SIRAS phasing while single wavelength anomalous dispersion (SAD) phasing was unsuccessful with more than 80,000 patterns of derivative crystals. We employed high energy X-rays from SACLA (12.6 keV) to take advantage of the large anomalous enhancement near the L III absorption edge of Hg, which is one of the most widely used heavy atoms for phasing in conventional protein crystallography. Hard XFEL is of benefit for de novo phasing in the use of routinely used heavy atoms and high resolution data collection.",
author = "Keitaro Yamashita and Dongqing Pan and Tomohiko Okuda and Michihiro Sugahara and Atsushi Kodan and Tomohiro Yamaguchi and Tomohiro Murai and Keiko Gomi and Naoki Kajiyama and Eiichi Mizohata and Mamoru Suzuki and Eriko Nango and Kensuke Tono and Yasumasa Joti and Takashi Kameshima and Jaehyun Park and Changyong Song and Takaki Hatsui and Makina Yabashi and So Iwata and Hiroaki Kato and Hideo Ago and Masaki Yamamoto and Toru Nakatsu",
note = "Funding Information: The authors are grateful to T. Nakane, O. Johnson, H. Powell, and A. Leslie for allowing us to use the unreleased version of MOSFLM with prior-cell function, which is now released as version 7.2.0, and to M. Maher and S. Hasnain for careful reading of the manuscript. The XFEL experiments were carried out at the BL3 of SACLA with the approval of the Japan Synchrotron Radiation Research Institute (JASRI) (proposal nos. 2014A8032 and 2013B8045). The SR experiment was carried out at the BL26B2 of SPring-8 with the approval of JASRI (proposal nos. 2014B1563, 2014A1845, and 2014A1858). This work was supported by the X-ray Free-Electron Laser Priority Strategy Program (MEXT) and JSPS KAKENHI Grant Number 60006047. The authors thank SACLA and SPring-8 BL26B2 beamline staff for technical assistance.",
year = "2015",
month = sep,
day = "11",
doi = "10.1038/srep14017",
language = "English",
volume = "5",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
}